Antifungal ethers

ABSTRACT

The present invention concerns compounds of formula                    
     the N-oxide forms, the salts, the quaternary amines and stereochemically isomeric forms thereof, wherein 
     D represents a tetrahydrofuran or dioxolane ring substituted with aryl and azolmethyl; —A—B— represents an optionally substituted bivalent radical of formula —N═CH—, —CH═N—, —CH═CH—, —CH 2 —CH 2 ; Alk represents C 1-6 alkanediyl; Y represents optionally substituted C 1-6 alkanediyl; R 1  and R 2  represent hydrogen, C 1-6 alkyl or arylC 1-6 alkyl; or R 1  and R 2  may be taken together to form an optionally substituted heterocyclic radical selected from morpholinyl, pyrrolidinyl, piperidinyl, homopiperidinyl, piperazinyl or phthalimid-1-yl; aryl represents phenyl, naphthalenyl, 1,2,3,4-tetrahydro-naphthalenyl, indenyl or indanyl; each of said aryl groups may optionally be substituted; having broad-spectrum antifungal activity; their preparation, compositions containing them and their use as a medicine.

This application is a National Stage application under 35 U.S.C. §371 ofPCT/EP00/03740 filed Apr. 20, 2000, which claims priority fromprovisional application 60/132,699, filed May 4, 1999.

The present invention is concerned with water soluble azole containingethers as broad-spectrum antifungals and their preparation; it furtherrelates to compositions comprising them, as well as their use as amedicine.

Systemic fungal infections in man are relatively rare in temperatecountries and many of the fungi that can become pathogenic normally livecommensally in the body or are common in the environment. The past fewdecades have witnessed an increasing incidence of numerouslife-threatening systemic fungal infections world-wide and these nowrepresent a major threat to many susceptible patients, particularlythose already hospitalized. Most of the increase can be attributed toimproved survival of immuno-compromised patients and the chronic use ofantimicrobial agents. Moreover, the flora typical of many common fungalinfections is also changing and this is presenting an epidemiologicalchallenge of increasing importance. Patients at greatest risk includethose with impaired immune functioning, either directly as a result ofimmunosuppression from cytotoxic drugs or HIV infection, or secondary toother debilitating diseases such as cancer, acute leukaemia, invasivesurgical techniques or prolonged exposure to antimicrobial agents. Themost common systemic fungal infections in man are candidosis,aspergillosis, histoplasmosis, coccidioidomycosis,paracoccidioidomycosis, blastomycosis and cryptococcosis.

Antifungals such as ketoconazole, itraconazole and fluconazole areemployed for the treatment and prophylaxis of systemic fungal infectionsin immuno-compromised patients. However, concern is growing about fungalresistance to some of these agents, especially these with a relativelynarrow spectrum, e.g. fluconazole. Worse still, it is recognized in themedical world that about 40% of the people suffering from severesystemic fungal infections are hardly, or not at all, able to receivemedication via oral administration. This inability is due to the factthat such patients are in coma or suffer from severe gastroparesis.Hence, the use of insoluble or sparingly soluble antifungals such asitraconazole, that are difficult to administer intravenously, is heavilyimpeded in this group of patients.

Also the treatment of onychomycosis, i.e. fungal infection of the nails,may well be served by potent water soluble antifungals. It is longdesired to treat onychomycosis via the transungual route. The problemthat then arises is to ensure that the antifungal agents will penetrateinto and beneath the nail. Mertin and Lippold (J. Pharm.

Pharmacol. (1997), 49, 30-34) stated that in order to screen for drugsfor topical application to the nail plate, attention has to be paidmainly to the water solubility of the compound. The maximum flux throughthe nail is beneficially influenced by increasing the water solubilityof the antifungal. Of course, efficacy in treating onychomycosis via thetransungual route is also dependent on the potency of the antifungal.

Consequently, there is a need for new antifungals, preferablybroad-spectrum antifungals, against which there is no existingresistance and which can be administered intravenously or transungually.Preferably the antifungal should also be available in a pharmaceuticalcomposition suitable for oral administration. This enables the physicianto continue treatment with the same drug after the patient has recoveredfrom the condition which required intravenous or transungualadministration of said drug.

U.S. Pat. No. 4,267,179 discloses heterocyclic derivatives of(4-phenylpiperazin-1-yl-aryloxy-methyl-1,3-dioxolan-2-yl)-methyl-1H-imidazolesand 1H-1,2,4-triazoles useful as antifungal agents. Said patentencompasses itraconazole, which is available as a broad-spectrumantifungal on a world-wide basis.

EP-A-0, 118,138 and EP-A-0,228,125 disclose4-[4-[4-[4-[[2-aryl-2-azolyl-1,3dioxolan-4-yl]methoxy]phenyl]-1-piperazinyl]phenyl]-2,4-dihydro-2-alkyloxyalkyl-3H-1,2,4-triazol-3-onederivatives as antifungals. WO 95/17407 discloses tetrahydrofuranantifungals as well as WO 96/38443 and WO 97/00255. The latter twopublications disclose tetrahydrofuran antifungals, which are taught tobe soluble and/or suspendible in an aqueous medium suitable forintravenous administration, containing substitution groups readilyconvertible in vivo into hydroxy groups.

Unexpectedly, the compounds of the present invention are potentbroad-spectrum antifungals with good water solubility.

The present invention concerns compounds of formula

the N-oxide forms, the salts, the quaternary amines and stereochemicallyisomeric forms thereof, wherein

D represents a radical of formula

 wherein the dotted line represents the bond attaching D to theremainder of the compound of formula (1);

X is N or CH;

R³ is hydrogen or halo;

R⁴ is halo;

—A—B— represents a bivalent radical of formula

—N═CH—  (i),

—CH═N—  (ii),

—CH═CH—  (iii),

—CH₂—CH₂  (iv),

wherein one hydrogen atom in the radicals (i) and (ii) may be replacedby a C₁₋₄alkyl radical and one or more hydrogen atoms in radicals (iii)and (iv) may be replaced by a C₁₋₄alkyl radical;

Alk represents C₁₋₆alkanediyl;

Y represents C₁₋₆alkanediyl optionally substituted with one or twosubstituents selected from halo, hydroxy, mercapto, C₁₋₄alkyloxy,C₁₋₄alkylthio, aryloxy, arylthio, arylC₁₋₄alkyloxy, arylC₁₋₄alkylthio,cyano, amino, mono- or di(C₁₋₄alkyl)amino, mono- or di(aryl)amino, mono-or di(arylC₁₋₄alkyl)amino, C₁₋₄alkyloxycarbonylamino,benzyloxycarbonylamino, aminocarbonyl, carboxyl, C₁₋₄alkyloxycarbonyl,guanidinyl, aryl and Het;

R¹ represents hydrogen, C₁₋₄alkyl or arylC₁₋₆alkyl; R² representshydrogen, C₁₋₄alkyl or arylC₁₋₆alkyl; or R¹ and R² may be taken togetherto form a heterocyclic radical selected from morpholinyl, pyrrolidinyl,piperidinyl, homopiperidinyl, piperazinyl or phthalimid-1-yl; saidheterocyclic radical may optionally be substituted with C₁₋₄alkyl, aryl,Het, arylC₁₋₄alkyl, HetC₁₋₄alkyl, hydroxyC₁₋₄alkyl, amino, mono- ordi(C₁₋₄alkyl)amino, aminoC₁₋₄alkyl, mono- ordi(C₁₋₄alkyl)aminoC₁₋₄alkyl, carboxyl, aminocarbonyl,C₁₋₄alkyloxycarbonyl; C₁₋₄alkyloxycarbonylamino or mono- ordi(C₁₋₄alkyl)aminocarbonyl;

aryl represents phenyl, naphthalenyl, 1,2,3,4-tetrahydro-naphthalenyl,indenyl or indanyl; each of said aryl groups may optionally besubstituted with one or more substituents selected from halo, C₁₋₄alkyl,hydroxy, C₁₋₄alkyloxy, nitro, amino, trifluoromethyl, hydroxyC₁₋₄alkyl,C₁₋₄alkyloxyC₁₋₄alkyl, aminoC₁₋₄alkyl, mono- ordi(C₁₋₄alkyl)aminoC₁₋₄alkyl;

Het represents a monocyclic or bicyclic heterocyclic radical; saidmonocyclic heterocyclic radical being selected from the grouppiperazinyl, homopiperazinyl, 1,4-dioxanyl, morpholinyl,thiomorpholinyl, pyridyl, piperidinyl, homopiperidinyl, pyrazinyl,pyrimidinyl, pyridazinyl, triazinyl, triazolyl, pyranyl,tetrahydropyranyl, imidazolyl, imidazolinyl, imidazolidinyl, pyrazolyl,pyrazolinyl, pyrazolidinyl, thiazolyl, thiazolidinyl, isothiazolyl,oxazolyl, oxazolidinyl, isoxazolyl, pyrrolyl, pyrrolinyl, pyrrolidinyl,furanyl, tetrahydrofuranyl, thienyl, thiolanyl, dioxolanyl; saidbicyclic heterocyclic radical being selected from the group quinolinyl,1,2,3,4-tetrahydroquinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl,phtalazinyl, cinnolinyl, chromanyl, thiochromanyl, 2H-chromenyl,1,4-benzodioxanyl, indolyl, isoindolyl, indolinyl, indazolyl, purinyl,pyrrolopyridinyl, furanopyridinyl, thienopyridinyl, benzothiazolyl,benzoxazolyl, benzisothiazolyl, benzisoxazolyl, benzimidazolyl,benzofuranyl, benzothienyl; whereby each of said mono- or bicyclicheterocycle may optionally be substituted with one or where possiblemore substituents selected from halo, C₁₋₄alkyl, hydroxy, C₁₋₄alkyloxy,nitro, amino, trifluoromethyl, hydroxyC₁₋₄alkyl, C₁₋₄alkyloxy-C₁₋₄alkyl,aminoC₁₋₄alkyl, mono- or di(C₁₋₄alkyl)aminoC₁₋₄alkyl, aryl orarylC₁₋₄alkyl.

As used in the foregoing definitions and hereinafter halo definesfluoro, chloro, bromo and iodo; C₁₋₄alkyl as a group or part of a groupencompasses the straight and branched chain saturated hydrocarbonradicals having from 1 to 4 carbon atoms such as, for example, methyl,ethyl, propyl, butyl and the like; C₁₋₆alkyl as a group or part of agroup encompasses the straight and branched chain saturated hydrocarbonradicals as defined in C₁₋₄alkyl as well as the higher homologuesthereof containing 5 or 6 carbon atoms such as, for example, pentyl orhexyl; C₁₋₆alkanediyl encompasses the straight and branched chainsaturated bivalent hydrocarbon radicals having from 1 to 6 carbon atomssuch as, for example, methylene, 1,2-ethanediyl, 1,3-propanediyl,1,4-butanediyl, 1,5-pentanediyl, 1,6-hexanediyl, 1,2-propanediyl,1,2-butanediyl, 2,3-butanediyl and the like; C₂₋₃alkanediyl encompassesthe straight and branched chain saturated bivalent hydrocarbon radicalshaving 2 or 3 carbon atoms such as, for example, 1,2-ethanediyl,1,2-propanediyl and 1,3-propanediyl.

For therapeutic use, salts of the compounds of formula (I) are thosewherein the counterion is pharmaceutically acceptable. However, salts ofacids and bases which are non-pharmaceutically acceptable may also finduse, for example, in the preparation or purification of apharmaceutically acceptable compound. All salts, whetherpharmaceutically acceptable or not are included within the ambit of thepresent invention.

The pharmaceutically acceptable addition salts as mentioned hereinaboveare meant to comprise the therapeutically active non-toxic acid additionsalt forms which the compounds of formula (I) are able to form. Thelatter can conveniently be obtained by treating the base form with suchappropriate acids as inorganic acids, for example, hydrohalic acids,e.g. hydrochloric, hydrobromic and the like; sulfuric acid; nitric acid;phosphoric acid and the like; or organic acids, for example, acetic,propanoic, hydroxyacetic, 2-hydroxypropanoic, 2-oxopropanoic, oxalic,malonic, succinic, maleic, fumaric, malic, tartaric,2-hydroxy-1,2,3-propanetricarboxylic, methanesulfonic, ethanesulfonic,benzenesulfonic, 4-methylbenzenesulfonic, cyclohexanesulfamic,2-hydroxybenzoic, 4-amino-2-hydroxybenzoic and the like acids.Conversely the salt form can be converted by treatment with alkali intothe free base form.

The compounds of formula (I) containing acidic protons may be convertedinto their therapeutically active non-toxic metal or amine addition saltforms by treatment with appropriate organic and inorganic bases.Appropriate base salt forms comprise, for example, the ammonium salts,the alkali and earth alkaline metal salts, e.g. the lithium, sodium,potassium, magnesium, calcium salts and the like, salts with organicbases, e.g. primary, secondary and tertiary aliphatic and aromaticamines such as methylamine, ethylamine, propylamine, isopropylamine, thefour butylamine isomers, dimethylamine, diethylamine, diethanolamine,dipropylamine, diisopropylamine, di-n-butylamine, pyrrolidine,piperidine, morpholine, trimethylamine, triethylamine, tripropylamine,quinuclidine, pyridine, quinoline and isoquinoline, the benzathine,N-methyl-D-glucamine, 2-amino-2-(hydroxymethyl)-1,3-propanediol,hydrabamine salts, and salts with amino acids such as, for example,arginine, lysine and the like. Conversely the salt form can be convertedby treatment with acid into the free acid form. The term addition saltalso comprises the hydrates and solvent addition forms which thecompounds of formula (I) are able to form. Examples of such forms aree.g. hydrates, alcoholates and the like.

The term “quaternary amine” as used hereinbefore defines the quaternaryammonium salts which the compounds of formula (I) are able to form byreaction between a basic nitrogen of a compound of formula (I) and anappropriate quaternizing agent, such as, for example, an optionallysubstituted alkylhalide, arylhalide or arylalkylhalide, e.g.methyliodide or benzyliodide. Other reactants with good leaving groupsmay also be used, such as alkyl trifluoromethanesulfonates, alkylmethanesulfonates, and alkyl p-toluenesulfonates. A quaternary amine hasa positively charged nitrogen. Pharmaceutically acceptable counterionsinclude chloro, bromo, iodo, trifluoroacetate and acetate. Thecounterion of choice can be made using ion exchange resin columns.

The term “stereochemically isomeric forms” as used hereinbefore definesall the possible stereoisomeric forms in which the compounds of formula(I) exist, thus, also including all enantiomers, enantiomeric mixturesand diastereomeric mixtures. Unless otherwise mentioned or indicated,the chemical designation of compounds denotes the mixture of allpossible stereoisomeric forms, said mixtures containing alldiastereomers and enantiomers of the basic molecular structure. The sameapplies to the intermediates as described herein, used to prepare endproducts of formula (I).

Pure stereoisomeric forms of the compounds and intermediates asmentioned herein are defined as isomers substantially free of otherenantiomeric or diastereomeric forms of the same basic molecularstructure of said compounds or intermediates. In particular, the term‘stereoisomerically pure’ being equivalent to ‘chirally pure’ concernscompounds or intermediates having a stereoisomeric excess of at least80% (i.e. minimum 90% of one isomer and maximum 10% of the otherpossible isomers) up to a stereoisomeric excess of 100% (i.e. 100% ofone isomer and none of the other), more in particular, compounds orintermediates having a stereoisomeric excess of 90% up to 100%, evenmore in particular having a stereoisomeric excess of 94% up to 100% andmost in particular having a stereoisomeric excess of 97% up to 100%. Theterms ‘enantiomerically pure’ and ‘diastereomerically pure’ should beunderstood in a similar way, but then having regard to the enantiomericexcess, respectively the diastereomeric excess of the mixture inquestion.

The terms cis and trans are used herein in accordance with ChemicalAbstracts nomenclature and refer to the position of the substituents ona ring moiety, more in particular on the tetrahydrofuran ring or thedioxolane ring in the compounds of formula (I). For instance, whenestablishing the cis or trains configuration of a tetrahydrofuran ordioxolane ring in a radical of formula (D₁), (D₂), (D₃) or (D₄), thesubstituent with the highest priority on the carbon atom in the 2position of the tetrahydrofuran or dioxolane ring, and the substituentwith the highest priority on the carbon atom in the 4 position of thetetrahydrofuran or dioxolane ring in the radicals of formula (D₁), (D2)or (D₄) or the 5 position of the tetrahydrofuran ring in the radical offormula (D₃) are considered (the priority of a substituent beingdetermined according to the Cahn-Ingold-Prelog sequence rules). Whensaid two substituents with highest priority are at the same side of thering then the configuration is designated cis, if not, the configurationis designated trans.

The compounds of formula (I) all contain at least 2 asymmetric centerswhich may have the R- or S-configuration. As used herein, thestereochemical descriptors denoting the stereochemical configuration ofeach of the 2 or more asymmetric centers are also in accordance withChemical Abstracts nomenclature.

Of some compounds of formula (I) and of intermediates used in theirpreparation, the absolute stereochemical configuration was notexperimentally determined. In those cases the stereoisomeric form whichwas first isolated is designated as “A” and the second as “B”, withoutfurther reference to the actual stereochemical configuration. However,said “A” and “B” stereoisomeric forms can be unambiguously characterizedby for instance their optical rotation in case “A” and “B” have anenantiomeric relationship. A person skilled in the art is able todetermine the absolute configuration of such compounds using art-knownmethods such as, for example, X-ray diffraction. In case “A” and “B” arestereoisomeric mixtures, they can be further separated whereby therespective first fractions isolated are designated “A1” and “B1” and thesecond as “A2” and “B2”, without further reference to the actualstereochemical configuration.

The N-oxide forms of the present compounds are meant to comprise thecompounds of formula (I) wherein one or several nitrogen atoms areoxidized to the so-called N-oxide.

Some of the compounds of formula (I) may also exist in their tautomericform. Such forms although not explicitly indicated in the above formulaare intended to be included within the scope of the present invention.

Whenever used hereinafter, the term “compounds of formula (I)” is meantto also include their N-oxide forms, their salts, their quaternaryamines and their stereochemically isomeric forms. Of special interestarc those compounds of formula (I) which are stereochemically pure.

An interesting group of compounds are those compounds of formula (I)wherein —Alk— is a bivalent radical of formula

wherein the carbon atom marked with one asterisk is attached to thenitrogen atom and the carbon atom marked with two asterisks is attachedto the oxygen atom.

Also interesting are those compounds of formula (I) wherein D is aradical of formula D₁or D₂, in particular D₁. Suitably, R³ and R⁴ areboth a halogen, more in particular a chloro or fluoro atom, and X is anitrogen atom.

Further —A—B— suitably is a radical of formula (ii).

A particular group of compounds are those compounds of formula (I)wherein Y is C₁₋₆alkanediyl optionally substituted with aryl; more inparticular, C₂₋₃alkanediyl optionally substituted with aryl.

A preferred group of compounds are those compounds of formula (1)wherein D is a radical of formula D₁, X is a nitrogen atom, R³ and R⁴are both a halogen, —Alk— is a bivalent radical of formula

wherein the carbon atom marked with an asterisk is attached to thenitrogen atom and the carbon atom marked with two asterisks is attachedto the oxygen atom and Y is C₂₋₃alkanediyl optionally substituted witharyl.

The compounds of the present invention can be prepared by reacting anintermediate of formula (II) wherein W¹ is a suitable leaving group suchas, for example, a halogen, e.g. iodo, an arylsulfonyloxy or analkanesulfonyloxy group, e.g. p-toluenesulfonyloxy, naphthylsulfonyloxyor methanesulfonyloxy, with an intermediate of formula (III) in areaction-inert solvent such as, for example, N,N-dimethylformamide,N,N-dimethylacetamide, 1-methyl-2-pyrrolidinone,1,3-dimethyl-2-imidazolidinone, sulfolane or the like, and in thepresence of a suitable base such as, for example, sodium hydroxide orsodium hydride.

In this and the following preparations, the reaction products may beisolated from the reaction medium and, if necessary, further purifiedaccording to methodologies generally known in the art such as, forexample, extraction, crystallization, trituration and chromatography. Inparticular, stereoisomers can be isolated chromatographically using achiral stationary phase such as, for example, Chiralpak AD (amylose 3,5dimethylphenyl carbamate) or Chiralpak AS, both purchased from DaicelChemical Industries, Ltd, in Japan.

Compounds of formula (I) may also be prepared by N-alkylating anintermediate of formula (IV) with an intermediate of formula (V) whereinW² is a suitable leaving group such as, for example, a halogen, and incase R¹ and/or R is hydrogen, the primary or secondary amine group isprotected with a protective group P such as, for example, aC₁₋₄alkyloxycarbonyl group or a benzyl group, in a reaction-inertsolvent such as, for example, dimethylsulfoxide, in the presence of abase such as, for example, potassium hydroxide. In case the amine wasprotected, art-known deprotection techniques can be employed to arriveat compounds of formula (I) after the N-alkylation reaction.

Compounds of formula (I) may also be prepared by reacting anintermediate of formula (VI) with an intermediate of formula (VII)wherein W3 is a suitable leaving group such as, for example, a halogen,an arylsulfonyloxy or an alkanesulfonyloxy group, e.g.p-toluenesulfonyloxy, naphthylsulfonyloxy or methanesulfonyloxy,optionally in the presence of a suitable base such as, for example,sodium hydride, and optionally in a reaction-inert solvent such as, forexample, N,N-dimethylformamide, N,N-dimethylacetamide, toluene,1-methyl-2-pyrrolidinone, 1,3-dimethyl-2-imidazolidinone, sulfolane orthe like. In case R¹ and/or R² is hydrogen, the primary or secondaryamine group may be protected with a protective group P such as, forexample, a C₁₋₄alkyloxycarbonyl group or a benzyl group, and after theO-alkylation reaction, art-known deprotection techniques can be employedto arrive at compounds of formula (I)

The compounds of formula (I) may also be converted into each otherfollowing art-known transformations.

Compounds of formula (I) containing a C₁₋₄alkyloxycarbonylamino moietymay be converted to compounds of formula (I) containing thecorresponding amino moiety using art-known techniques such as, forexample, reaction in dichloromethane and in the presence oftrifluoroacetic acid.

Compounds of formula (I) containing a primary amine may bemono-methylated by first protecting the primary amine with a suitableprotecting group such as, for example, an arylalkyl group, e.g. benzyl;and subsequently methylating the secondary amine using art-knownmethylation techniques such as, for example, reaction withparaformaldehyde. The thus obtained tertiary amine may be deprotectedusing art-known deprotection techniques such as, for example, reactionwith hydrogen in tetrahydrofuran or methanol and in the presence of acatalyst such as, for example palladium-on-charcoal, thus obtaining thedesired methylated secondary amine.

The compounds of formula (I) may also be converted to the correspondingN-oxide forms following art-known procedures for converting a trivalentnitrogen into its N-oxide form. Said N-oxidation reaction may generallybe carried out by reacting the starting material of formula (I) with anappropriate organic or inorganic peroxide. Appropriate inorganicperoxides comprise, for example, hydrogen peroxide, alkali metal orearth alkaline metal peroxides, e.g. sodium peroxide, potassiumperoxide; appropriate organic peroxides may comprise peroxy acids suchas, for example, benzenecarboperoxoic acid or halo substitutedbenzenecarboperoxoic acid, e.g. 3-chlorobenzenecarboperoxoic acid,peroxoalkanoic acids, e.g. peroxoacetic acid, alkylhydroperoxides, e.g.tert-butyl hydroperoxide. Suitable solvents are, for example, water,lower alkanols, e.g. ethanol and the like, hydrocarbons, e.g. toluene,ketones, e.g. 2-butanone, halogenated hydrocarbons, e.g.dichloromethane, and mixtures of such solvents.

Some of the intermediates and starting materials used in the abovereaction procedures are commercially available, or may be synthesizedaccording to procedures described elsewhere, e.g. U.S. Pat. No.4,619,931, U.S. Pat. No. 4,791,111, U.S. Pat. No. 4,931,444, U.S. Pat.No. 4,267,179 and WO 98/34934.

Pure stereoisomeric forms of the compounds and the intermediates of thisinvention may be obtained by the application of art-known procedures.Diastereomers may be separated by physical separation methods such asselective crystallization and chromatographic techniques, e.g. liquidchromatography using chiral stationary phases.

Enantiomers may be separated from each other by the selectivecrystallization of their diastereomeric salts with optically activeacids. Alternatively, enantiomers may be separated by chromato-graphictechniques using chiral stationary phases. Said pure stereoisomericforms may also be derived from the corresponding pure stereoisomericforms of the appropriate starting materials, provided that the reactionoccurs stereo-selectively or stereospecifically. Preferably if aspecific stereoisomer is desired, said compound will be synthesized bystereoselective or stereospecific methods of preparation. These methodswill advantageously employ chirally pure starting materials.Stereoisomeric forms of the compounds of formula (1) are obviouslyintended to be included within the scope of the invention.

The chirally pure forms of the compounds of formula (I) form a preferredgroup of compounds. It is therefore that the chirally pure forms of theintermediates of formula (II), (III) and (VI), their N-oxide forms,their salt forms and their quaternary amines are particularly useful inthe preparation of chirally pure compounds of formula (I). Alsoenantiomenc mixtures and diastereomeric mixtures of intermediates offormula (II), (III) and (VI) are useful in the preparation of compoundsof formula (I) with the corresponding configuration.

The compounds of formula (I), the salts, the quaternary amines and thestereochemically isomeric forms thereof are useful agents for combatingfungi in vivo. The present compounds are broad-spectrum antifungals.They are active against a wide variety of fungi, such as Candida spp.,e.g. Candida albicans, Candida glabrata, Candida krusei, Candidaparapsilosis, Candida kefyr, Candida tropicalis; Aspergillus spp., e.g.Aspergillus fumigatus, Aspergillus niger, Aspergillus flavus;Cryptococcus neoformans; Sporothrix schenckii; Fonsecaea spp.;Epidermophyton floccosum; Microsporum canis; Trichophyton spp.; Fusariumspp.; and several dematiaceous hyphomycetes. Of particular interest isthe improved activity of some of the present compounds against Fusariumspp.

In vitro experiments, including the determination of the fungalsusceptibility of the present compounds as described in thepharmacological example hereinafter, indicate that the compounds offormula (I) have a favourable intrinsic inhibitory capacity on fungalgrowth in for instance Candida albicans. Other in vitro experiments suchas the determination of the effects of the present compounds on thesterol synthesis in, for instance, Candida albicans, also demonstratetheir antifungal potency. Also in vivo experiments in several mouse,guinea-pig and rat models show that, after both oral and intravenousadministration, the present compounds are potent antifungals.

An additional advantage of some of the present compounds is that theyare not only fungistatic, as most of the known azole antifungals, butare also fungicidal at acceptable therapeutic doses against many fungalisolates.

The compounds of the present invention are chemically stable and have agood oral availability.

The solubility profile in aqueous solutions of the compounds of formula(I) makes them suitable for intravenous administration. Particularlyinteresting compounds are those compounds of formula (I) having awater-solubility of at least 0.1 mg/ml at a pH of at least 4,preferably, a water-solubility of at least 1 mg/ml at a pH of at least4, and more preferred a water-solubility of at least 5 mg/ml at a pH ofat least 4.

In view of the utility of the compounds of formula (I), there isprovided a method of treating warm-blooded animals, including humans,suffering from fungal infections. Said method comprises the systemic ortopical administration of an effective amount of a compound of formula(I), a N-oxide form, a salt, a quaternary amine or a possiblestereoisomeric form thereof, to warm-blooded animals, including humans.Hence, compounds of formula (I) are provided for use as a medicine, inparticular, the use of a compound of formula (I) in the manufacture of amedicament useful in treating fungal infections is provided.

The present invention also provides compositions for treating orpreventing fungal infections comprising a therapeutically effectiveamount of a compound of formula (I) and a pharmaceutically acceptablecarrier or diluent.

In view of their useful pharmacological properties, the subjectcompounds may be formulated into various pharmaceutical forms foradministration purposes. To prepare the pharmaceutical compositions ofthis invention, a therapeutically effective amount of a particularcompound, in base or addition salt form, as the active ingredient iscombined in intimate admixture with a pharmaceutically acceptablecarrier, which.carrier may take a wide variety of forms depending on theform of preparation desired for administration. These pharmaceuticalcompositions are desirably in unitary dosage form suitable, preferably,for administration orally, rectally, topically, percutaneously,transungually or by parenteral injection. For example, in preparing thecompositions in oral dosage form, any of the usual pharmaceutical mediamay be employed, such as, for example, water, glycols, oils, alcoholsand the like in the case of oral liquid preparations such assuspensions, syrups, emulsions, elixirs and solutions: or solid carrierssuch as starches, sugars, kaolin, lubricants, binders, disintegratingagents and the like in the case of powders, pills, capsules and tablets.Because of their ease in administration, tablets and capsules representthe most advantageous oral dosage unit form, in which case solidpharmaceutical carriers are obviously employed. As appropriatecompositions for topical application there may be cited all compositionsusually employed for topically administering drugs e.g. creams, gel,dressings, shampoos, tinctures, pastes, ointments, salves, powders andthe like. In the compositions suitable for percutaneous administration,the carrier optionally comprises a penetration enhancing agent and/or asuitable wetting agent, optionally combined with suitable additives ofany nature in minor proportions, which additives do not cause asignificant deleterious effect to the skin. Said additives mayfacilitate the administration to the skin and/or may be helpful forpreparing the desired compositions. These compositions may beadministered in various ways, e.g., as a transdermal patch, as aspot-on, as an ointment.

Transungual compositions are in the form of a solution and the carrieroptionally comprises a penetration enhancing agent which favours thepenetration of the antifungal into and through the keratinized unguallayer of the nail. The solvent medium comprises water mixed with aco-solvent such as an alcohol having from 2 to 6 carbon atoms, e.g.ethanol.

For parenteral compositions, the carrier will usually comprise sterilewater, at least in large part. Injectable solutions, for example, may beprepared in which the carrier comprises saline solution, glucosesolution or a mixture of saline and glucose solution. Injectablesuspensions may also be prepared in which case appropriate liquidcarriers, suspending agents and the like may be employed. For parenteralcompositions, also other ingredients, to aid solubility for example,e.g. cyclodextrins, may be included. Appropriate cyclodextrins are α-,β-, γ-cyclodextrins or ethers and mixed ethers thereof wherein one ormore of the hydroxy groups of the anhydroglucose units of thecyclodextrin are substituted with C₁₋₆alkyl, particularly methyl, ethylor isopropyl, e.g. randomly methylated β-CD; hydroxyC₁₋₆alkyl,particularly hydroxyethyl, hydroxypropyl or hydroxybutyl;carboxyC₁₋₆alkyl, particularly carboxymethyl or carboxyethyl;C₁₋₆alkylcarbonyl, particularly acetyl. Especially noteworthy ascomplexants and/or solubilizers are β-CD, randomly methylated β-CD,2,6-dimethyl-β-CD, 2-hydroxyethyl-β-CD, 2-hydroxyethyl-γ-CD,2-hydroxypropyl-γ-CD and (2-carboxymethoxy)propyl-β-CD, and inparticular 2-hydroxypropyl-β-CD (2-HP-β-CD).

The term mixed ether denotes cyclodextrin derivatives wherein at leasttwo cyclodextrin hydroxy groups are etherified with different groupssuch as, for example, hydroxy-propyl and hydroxyethyl.

The average molar substitution (M.S.) is used as a measure of theaverage number of moles of alkoxy units per mole of anhydroglucose. Theaverage substitution degree (D.S.) refers to the average number ofsubstituted hydroxyls per anhydroglucose unit. The M.S. and D.S. valuecan be determined by various analytical techniques such as nuclearmagnetic resonance (NMR), mass spectrometry (MS) and infraredspectroscopy (IR). Depending on the technique used, slightly differentvalues may be obtained for one given cyclodextrin derivative.Preferably, as measured by mass spectrometry, the M.S. ranges from 0.125to 10 and the D.S. ranges from 0.125 to 3.

Other suitable compositions for oral or rectal administration compriseparticles consisting of a solid dispersion comprising a compound offormula (I) and one or more appropriate pharmaceutically acceptablewater-soluble polymers.

The term “a solid dispersion” used hereinafter defines a system in asolid state (as opposed to a liquid or gaseous state) comprising atleast two components, in casu the compound of formula (I) and thewater-soluble polymer, wherein one component is dispersed more or lessevenly throughout the other component or components (in case additionalpharmaceutically acceptable formulating agents, generally known in theart, are included, such as plasticizers, preservatives and the like).When said dispersion of the components is such that the system ischemically and physically uniform or homogenous throughout or consistsof one phase as defined in thermo-dynamics, such a solid dispersion willbe called “a solid solution”. Solid solutions are preferred physicalsystems because the components therein are usually readily bioavailableto the organisms to which they are administered. This advantage canprobably be explained by the ease with which said solid solutions canform liquid solutions when contacted with a liquid medium such as thegastro-intestinal juices. The ease of dissolution may be attributed atleast in part to the fact that the energy required for dissolution ofthe components from a solid solution is less than that required for thedissolution of components from a crystalline or microcrystalline solidphase.

The term “a solid dispersion” also comprises dispersions which are lesshomogenous throughout than solid solutions. Such dispersions are notchemically and physically uniform throughout or comprise more than onephase. For example, the term “a solid dispersion” also relates to asystem having domains or small regions wherein amorphous,microcrystalline or crystalline compound of formula (I), or amorphous,microcrystalline or crystalline water-soluble polymer, or both, aredispersed more or less evenly in another phase comprising water-solublepolymer, or compound of formula (I), or a solid solution comprisingcompound of formula (I) and water-soluble polymer. Said domains areregions within the solid dispersion distinctively marked by somephysical feature, small in size, and evenly and randomly distributedthroughout the solid dispersion.

Various techniques exist for preparing solid dispersions includingmelt-extrusion, spray-drying and solution-evaporation.

The solution-evaporation process comprises the following steps:

a) dissolving the compound of formula (I) and the water-soluble polymerin an appropriate solvent, optionally at elevated temperatures;

b) heating the solution resulting under point a), optionally undervacuum, until the solvent is evaporated. The solution may also be pouredonto a large surface so as to form a thin film, and evaporating thesolvent therefrom.

In the spray-drying technique, the two components are also dissolved inan appropriate solvent and the resulting solution is then sprayedthrough the nozzle of a spray dryer followed by evaporating the solventfrom the resulting droplets at elevated temperatures.

The preferred technique for preparing solid dispersions is themelt-extrusion process comprising the following steps:

a) mixing a compound of formula (I) and an appropriate water-solublepolymer,

b) optionally blending additives with the thus obtained mixture,

c) heating and compounding the thus obtained blend until one obtains ahomogenous melt,

d) forcing the thus obtained melt through one or more nozzles; and

e) cooling the melt till it solidifies.

The terms “melt” and “melting” should be interpreted broadly. Theseterms not only mean the alteration from a solid state to a liquid state,but can also refer to a transition to a glassy state or a rubbery state,and in which it is possible for one component of the mixture to getembedded more or less homogeneously into the other. In particular cases,one component will melt and the other component(s) will dissolve in themelt thus forming a solution, which upon cooling may form a solidsolution having advantageous dissolution properties.

After preparing the solid dispersions as described hereinabove, theobtained products can be optionally milled and sieved.

The solid dispersion product can be milled or ground to particles havinga particle size of less than 600 μm, preferably less than 400 μm andmost preferably less than 125 μm.

The particles prepared as described hereinabove can then be formulatedby conventional techniques into pharmaceutical dosage forms such astablets and capsules.

It will be appreciated that a person of skill in the art will be able tooptimize the parameters of the solid dispersion preparation techniquesdescribed above, such as the most appropriate solvent, the workingtemperature, the kind of apparatus being used, the rate of spray-drying,the throughput rate in the melt-extruder

The water-soluble polymers in the particles are polymers that have anapparent viscosity, when dissolved at 20° C. in an aqueous solution at2% (w/v), of 1 to 5000 mPa.s more preferably of 1 to 700 mPa.s, and mostpreferred of 1 to 100 mPa.s. For example, suitable water-solublepolymers include alkylcelluloses, hydroxyalkylcelluloses, hydroxyalkylalkylcelluloses, carboxyalkylcelluloses, alkali metal salts ofcarboxyalkylcelluloses, carboxyalkylalkylcelluloses,carboxyalkylcellulose esters, starches, pectines, chitin derivates, di-,oligo- and polysaccharides such as trehalose, alginic acid or alkalimetal and ammonium salts thereof, carrageenans, galactomannans,tragacanth, agar-agar, gummi arabicum, guar gummi and xanthan gummi,polyacrylic acids and the salts thereof, polymethacrylic acids and thesalts thereof, methacrylate copolymers, polyvinylalcohol,polyvinylpyrrolidone, copolymers of polyvinyl-pyrrolidone with vinylacetate, combinations of polyvinylalcohol and polyvinyl-pyrrolidone,polyalkylene oxides and copolymers of ethylene oxide and propyleneoxide. Preferred water-soluble polymers are hydroxypropylmethylcelluloses.

Also one or more cyclodextrins can be used as water soluble polymer inthe preparation of the above-mentioned particles as is disclosed in WO97/18839. Said cyclodextrins include the pharmaceutically acceptableunsubstituted and substituted cyclodextrins known in the art, moreparticularly α, β or γ cyclodextrins or the pharmaceutically acceptablederivatives thereof.

Substituted cyclodextrins which can be used include polyethers describedin U.S. Pat. No. 3,459,731. Further substituted cyclodextrins are etherswherein the hydrogen of one or more cyclodextrin hydroxy groups isreplaced by C₁₋₆alkyl, hydroxyC₁₋₆alkyl, carboxy-C₁₋₆alkyl orC₁₋₆alkyloxycarbonylC₁₋₆alkyl or mixed ethers thereof. In particularsuch substituted cyclodextrins are ethers wherein the hydrogen of one ormore cyclodextrin hydroxy groups is replaced by C₁₋₃alkyl,hydroxyC₂₋₄alkyl or carboxyC₁₋₂alkyl or more in particular by methyl,ethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, carboxy-methyl orcarboxyethyl.

As used hereinbefore, C₁₋₂alkyl represents straight or branched chainsaturated hydrocarbon radicals having 1 or 2 carbon atoms such as methylor ethyl; C₁₋₃alkyl encompasses the straight and branched chainsaturated hydrocarbon radicals as defined in C₁₋₂alkyl as well as thehigher homologue thereof containing 3 carbon atoms, such as propyl;C₂₋₄alkyl represents straight or branched chain saturated hydrocarbonradicals having from 2 to 4 carbon atoms such as ethyl, propyl, butyl,1-methyl-propyl and the like.

Of particular utility are the β-cyclodextrin ethers, e.g.dimethyl-β-cyclodextrin as described in Drugs of the Future, Vol. 9, No.8, p. 577-578 by M. Nogradi (1984) and polyethers, e.g. hydroxypropylβ-cyclodextrin and hydroxyethyl β-cyclodextrin, being examples. Such analkyl ether may be a methyl ether with a degree of substitution of about0.125 to 3, e.g. about 0.3 to 2. Such a hydroxypropyl cyclodextrin mayfor example be formed from the reaction between β-cyclodextrin anpropylene oxide and may have a MS value of about 0.125 to 10, e.g. about0.3 to 3.

Another suitable type of substituted cyclodextrins issulfobutylcyclodextrines.

The ratio of active ingredient over cyclodextrin may vary widely. Forexample ratios of 1/100 to 100/1 may be applied. Interesting ratios ofactive ingredient over cyclodextrin range from about 1/10 to 10/1. Moreinteresting ratios of active ingredient over cyclodextrin range fromabout 1/5 to 5/1.

It may further be convenient to formulate the present azole antifungalsin the form of nanoparticles which have a surface modifier adsorbed onthe surface thereof in an amount sufficient to maintain an effectiveaverage particle size of less than 1000 nm. Useful surface modifiers arebelieved to include those which physically adhere to the surface of theantifungal agent but do not chemically bond to the antifungal agent.

Suitable surface modifiers can preferably be selected from known organicand inorganic pharmaceutical excipients. Such excipients include variouspolymers, low molecular weight oligomers, natural products andsurfactants. Preferred surface modifiers include nonionic and anionicsurfactants.

Yet another interesting way of formulating the present compoundsinvolves a pharmaceutical composition whereby the present antifungalsare incorporated in hydrophilic polymers and applying this mixture as acoat film over many small beads, thus yielding a composition which canconveniently be manufactured and which is suitable for preparingpharmaceutical dosage forms for oral administration.

Said beads comprise a central, rounded or spherical core, a coating filmof a hydrophilic polymer and an antifungal agent and a seal-coatinglayer.

Materials suitable for use as cores in the beads are manifold, providedthat said materials are pharmaceutically acceptable and have appropriatedimensions and firmness. Examples of such materials are polymers,inorganic substances, organic substances, and saccharides andderivatives thereof.

The pharmaceutical compositions mentioned above may also contain afungicidally effective amount of other antifungal compounds such as cellwall active compounds. The term “cell wall active compound”, as usedherein, means any compound which interferes with the fungal cell wall.Appropriate antifungal compounds for use in combination with the presentcompounds include, but are not limited to, known azoles such asfluconazole, voriconazole, itraconazole, ketoconazole, miconazole, ER30346, SCH 56592; polyenes such as amphotericin B, nystatin or liposomaland lipid forms thereof, such as Abelcet, AmBisome and Amphocil; purineor pyrimidine nucleotide inhibitors such as flucytosine; polyoxins andnikkcomycins, in particular nikkomycin Z or nikkomycin K and otherswhich are described in U.S. Pat. No. -5,006,513 or other chitininhibitors; elongation factor inhibitors such as sordarin and analogsthereof; mannan inhibitors such as predamycin;bactericidal/permeability-inducing (BPI) protein products such as XMP.97or XMP.127; complex carbohydrate antifungal agents such as CAN-296;(1,3)-β-glucan synthase inhibitors including papulacandins, aculeacins,and echinocandins.

It is especially advantageous to formulate the aforementionedpharmaceutical compositions in unit dosage form for ease ofadministration and uniformity of dosage. Unit dosage form as used in thespecification and claims herein refers to physically discrete unitssuitable as unitary dosages, each unit containing a predeterminedquantity of active ingredient calculated to produce the desiredtherapeutic effect in association with the required pharmaceuticalcarrier. Examples of such unit dosage forms are tablets (includingscored or coated tablets), capsules, pills, suppositories, powderpackets, wafers, injectable solutions or suspensions, teaspoonfuls,tablespoonfuls and the like, and segregated multiples thereof.

Those of skill in treating warm-blooded animals suffering from diseasescaused by fungi could easily determine the therapeutically effectivedaily amount from the test results given herein. In general, it iscontemplated that a therapeutically effective daily amount would be from0.05 mg/kg to 20 mg/kg body weight.

EXPERIMENTAL PART

Hereinafter, “DMF” is defined as N,N-dimethylformamide and “DIPE” isdefined as diisopropylether.

A. PREPARATION OF THE INTERMEDIATES Example A1

a) A mixture of2,4-dihydro4-[4-[4-(4-methoxyphenyl)-1-piperazinyl]phenyl]-3H-1,2,4-triazol-3-one(0.05 mol), 2-bromo-butanoic acid ethyl ester (0.055 mol) and Na₂CO₃(0.15 mol) in 1-methyl-2-pyrrolidinone (250 m1) was stirred at 75° C.over 2-Bromobutanoic acid ethyl ester (0.015 mol) was added again. Themixture was stirred at 75° C. for 6 hours, at room temperature for 48hours, poured out into H₂O and stirred for 30 minutes. The precipitatewas filtered off and dissolved in CH₂Cl₂. The solution was filtered. Thefiltrate was dried, filtered and the solvent was evaporated. The residuewas triturated in diisopropylether and ethylacetate, filtered off anddried, yielding 10 g (43%) of (±)-ethylα-ethyl-4,5-dihydro-4-[4-[4-(4-methoxyphenyl)-1-piperazinyl]phenyl]-5-oxo- 1H-1,2,4-triazol-1-acetate (interm. 1).

b) A mixture of NaHSO₃ (1 g) in HBr 48% (250 ml) and a acetic acid/HBrmixture (250 ml) was stirred for 15 minutes. Intermediate 1 (0.022 mol)was added. The mixture was stirred and refluxed for 90 minutes. Thesolvent was evaporated. Toluene was added and evaporated. The residuewas dissolved in CH₃OH. The mixture was stirred on an ice bath. SOCl₂(24 g) was added dropwise. The mixture was stirred overnight. Thesolvent was evaporated. The residue was dissolved in CH₂Cl₂. The organicsolution was washed with a NaHCO₃ solution, dried, filtered and thesolvent was evaporated. The residue was triturated in DIPE, filtered offand dried, yielding 6.6 g (±)-methylα-ethyl4,5-dihydro4-[4-[4-(4-hydroxyphenyl)-1-piperazinyl]phenyl]-5-oxo-1H-1,2,4-triazol-1-acetate(interm. 2).

c) A mixture of(−)-(2S-cis)-2-(2,4-difluorophenyl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1,3-dioxolane-4-methanolmethanesulfonate(ester) (0.007 mol), intermediate (2) (0.0068 mol) andNaOH (0.008 mol) in DMF (100 ml) was stirred at 50° C. under N₂ flowovernight, then poured out into H₂O and stirred for 1 hour. Theprecipitate was filtered off and dissolved in CH₂Cl₂. The organic layerwas separated, dried, filtered and the solvent was evaporated. Theresidue was purified by column chromatography over silica gel (eluent:CH₂Cl₂/CH₃OH/hexane/ethyl acetate 48/2/20/30). The pure fractions werecollected and the solvent was evaporated. The residue was triturated inethyl acetate, filtered off and dried, yielding 1.4 g (29%) of methyl(2S-cis)-4-[4-[4-[4-[[2-(2,4-difluorophenyl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1,3-dioxolan-4-yl]methoxy]-phenyl]-1-piperazinyl]phenyl]-α-ethyl4,5-dihydro-5-oxo-1H-1,2,4-triazol-1-acetate(interm. 3a).

In an analogous way, intermediate 3b was prepared.

d) A mixture of intermediate (3a) (0.009 mol) and NaBH₄ (0.045 mol) indioxane (300 ml) and H₂O (100 ml) was stirred at room temperatureovernight. A saturated NH₄Cl solution (100 ml) was added. The mixturewas stirred for 3 hours. HCl (10 ml) was added. The mixture was stirredfor 48 hours, then neutralized with a Na₂CO₃ solution and extracted withCH₂Cl₂. The organic layer was separated, washed, dried, filtered and thesolvent was evaporated. The residue was purified by columnchromatography over silica gel (eluent: CH₂Cl₂/CH₃OH 96/4). The purefractions were collected and the solvent was evaporated. The residue wastriturated in DIPE, filtered off and dried, yielding 4.2 g (68%)(2S-cis)4-[4-[4-[4[[2-(2,4-difluorophenyl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1,3-dioxolan4-yl]methoxy]phenyl]-1-piperazinyl]phenyl]-2,4-dihydro-2-[1-(hydroxymethyl)propyl]-3H-1,2,4-triazol-3-one(interm. 4).

B. PREPARATION OF THE FINAL COMPOUNDS Example B1 Preparation of Compound5

A mixture of intermediate 4 (0.006 mol) and2-chloro-N,N-diethyl-ethanamine hydrochloride (0.009 mol) in DMF (100ml) was stirred at 50° C. under N₂ flow. NaH (0.018 mol) was added. Themixture was stirred at 50° C. under N₂ flow overnight, then poured outinto H₂O and stirred for 30 minutes. The precipitate was filtered offand dissolved in CH₂Cl₂. The organic solution was washed, dried,filtered and the solvent was evaporated. The residue was purified bycolumn chromatography over silica gel (eluent: CH₂Cl₂/(CH₃OH/NH₃) 98/2).The pure fractions were collected and the solvent was evaporated. Theresidue was triturated in DIPE, filtered off and dried. The residue waspurified by HPLC over silica gel (eluent: CH₂Cl₂/CH₃OH/(CH₃OH/NH₃)96/2/2). The pure fractions were collected and the solvent wasevaporated. The residue was triturated in DIPE, filtered off and dried,yielding 0.95 g of compound 5.

Example B2 Preparation of Compound 3

A mixture of compound 2 (see Table 1) (0.0062 mol), prepared accordingto the procedure described in example B1, in tetrahydrofuran (150ml) washydrogenated for 6 days with palladium-on-charcoal 10% (2 g) as acatalyst. After uptake of hydrogen (2 equiv), the catalyst was filteredoff and the filtrate was evaporated. The residue was purified oversilica gel on a glass filter (eluent: CH₂Cl₂/CH₃OH 98/2 and 97/3). Thepure fractions were collected and the solvent was evaporated. Theresidue was crystallized from 2-propanol. The precipitate was filteredoff and dried, yielding 1.37 g (27%) of compound 3.

Table 1 lists the compounds which were prepared analogous to example B1or B2.

TABLE 1

Comp. Ex. No. No. L stereochemistry; optical rotation and/or meltingpoint 1 B1

[2S-[2α,4α(R*,R*)]]; [α]₂₀ ^(D) = −24.42 at concentration of 24.98 mg/5ml methanol 2 B1

[2S-[2α,4α[(R*,R*)(R*)]]] 3 B2

[2S-[2α,4α[(R*,R*)(R*)]]]; [α]₂₀ ^(D) = −30.76 at concentration of 25.03mg/5 ml DMF; mp. 128.7° C. 4 B1

2S-cis; [α]₂₀ ^(D) = −12.15 at concentration of 25.11 mg/5 ml DMF 5 B1

2S-cis 6 B1

[2S-[2α,4α[(R*,R*)(R*)]]]; [α]₂₀ ^(D) = −47.62 at concentration of 26.04mg/5 ml DMF; mp. 85.5° C.

C. PHYSICOCHEMICAL EXAMPLE Example C1 Water Solubility

An excess of compound was added to water buffered with 0.1 M citric acidand 0.2 M Na₂HPO₄ in a ratio of 61.5/38.5 (pH=4). The mixture was shakenduring 1 day at room temperature. The precipitate was filtered off. Theconcentration of the compound was measured via UV spectrospcopy and isshown in Table 2.

TABLE 2 Comp.No. pH solubility in mg/ml 1 4.05 >10 3 3.65 >7.09 44.4 >9.8 5 4.05 >12

D. PHARMACOLOGICAL EXAMPLE Example D1 Determination of FungalSusceptibility.

A panel of Candida isolates plus single isolates of the dermatophytesMicrosporum canis, Trichophyton rubrum and Trichophyton mentagrophytes;Aspergillus fumigatus, and Cryptococcus neoformans were used to evaluatethe activity of the test compounds in vitro. Inocula were prepared asbroth cultures (yeasts) or as suspensions of fungal material made fromagar slope cultures (moulds). The test compounds were pipetted fromdimethylsulfoxide stock solution into water to provide a series of10-fold dilutions. The fungal inocula were suspended in the growthmedium CYG (F. C. Odds, Journal of Clinical Microbiology, 29, 2735-2740,1991) at approximately 50,000 colony-forming units (CFU) per ml andadded to the aqueous test drugs. The cultures were set up in the 96wells of plastic microdilution plates and they were incubated for 2 daysat 37° C. (Candida spp.) or for 5 days at 30° C. (other fungi). Growthin the microcultures was measured by their optical density (OD) measuredat a wavelength of 405 nm. The OD for cultures with test compounds wascalculated as a percentage of the OD for control cultures, i.e. the ODfor cultures without test compounds. Inhibition of growth to 35% ofcontrol or less was recorded as significant inhibition.

Minimal inhibitory concentration (MIC; in 10⁻⁶ M) of some of thecompounds of formula (I) for Candida glabrata, Candida krusei, Candidaparapsilosis, Candida albicans, Candida kefyr, Candida tropicalis,Microsporum canis, Trichophyton rubrum, Trichophyton mentagrophytes,Cryptococcus neoformans and Aspergillus fumigatus are listed in table 3.

TABLE 3 MIC values in 10⁻⁶ M Infection Comp. 1 Comp. 3 Comp. 5 Candidaalbicans 0.1 <0.01 <0.1 Candida glabrata 10 1 10 Candida krusei 10 1 10Candida parapsilosis 0.1 <0.01 <0.1 Candida kefyr 0.1 <0.01 <0.1 Candidatropicalis 0.1 0.1 1 Microsporum canis 10 1 <0.1 Trichophyton rubrum 10.1 10 Trichophyton mentagrophytes 10 1 10 Cryptococcus neoformans 1 0.11 Aspergillus fumigatus 10 1 1

E. COMPOSITION EXAMPLE

“Active ingredient” (A.I.) as used throughout these examples relates toa compound of formula (I), a N-oxide, a salt, a quaternary amine or astereochemically isomeric form thereof.

Example E1 Injectable Solution.

1.8 Grams methyl 4-hydroxybenzoate and 0.2 grams sodium hydroxide weredissolved in about 0.5 l of boiling water for injection. After coolingto about 50° C. there were added while stirring 0.05 grams propyleneglycol and 4 grams of the active ingredient. The solution was cooled toroom temperature and supplemented with water for injection q.s. ad 1 l,giving a solution comprising 4 mg/ml of active ingredient. The solutionwas sterilized by filtration and tilled in sterile containers.

Example E2 Transungual Composition

0.144 g KH₂PO₄, 9 g NaCl, 0.528 g Na₂HPO₄.2H₂O was added to 800 ml H₂Oand the mixture was stirred. The pH was adjusted to 7.4 with NaOH and500 mg NaN₃ was added. Ethanol (42 v/v %) was added and the pH wasadjusted to 2.3 with HCl. 15 mg active ingredient was added to 2.25 mlPBS (Phosphate Buffer Saline)/Ethanol (42%; pH 2.3) and the mixture wasstirred and treated with ultrasound. 0.25 ml PBS/Ethanol (42%; pH2.3)was added and the mixture was further stirred and treated withultrasound until all active ingredient was dissolved, yielding thedesired transungual composition.

Example E3 Oral Drops

500 Grams of the A.I. was dissolved in 0.5 l of a sodium hydroxidesolution and 1.5 l of the polyethylene glycol at 60˜80° C. After coolingto 30˜40° C. there were added 35 l of polyethylene glycol and themixture was stirred well. Then there was added a solution of 1750 gramsof sodium saccharin in 2.5 l of purified water and while stirring therewere added 2.5 l of cocoa flavor and polyethylene glycol q.s. to avolume of 50 l, providing an oral drop solution comprising 10 mg/ml ofA.I. The resulting solution was filled into suitable containers.

Example E4 Capsules

20 Grams of the A.I., 6 grams sodium lauryl sulfate, 56 grams starch, 56grams lactose, 0.8 grams colloidal silicon dioxide, and 1.2 gramsmagnesium stearate were vigorously stirred together. The resultingmixture was subsequently filled into 1000 suitable hardened gelatincapsules, comprising each 20 mg of the active ingredient.

Example E5 Film-coated Tablets

Preparation of Tablet Core

A mixture of 100 grams of the A.I., 570 grams lactose and 200 gramsstarch was mixed well and thereafter humidified with a solution of 5grams sodium dodecyl sulfate and 10 grams polyvinylpyrrolidone in about200 ml of water. The wet powder mixture was sieved, dried and sievedagain. Then there was added 100 grams microcrystalline cellulose and 15grams hydrogenated vegetable oil. The whole was mixed well andcompressed into tablets, giving 10,000 tablets, each containing 10 mg ofthe active ingredient.

To a solution of 10 grams methyl cellulose in 75 ml of denaturatedethanol there was added a solution of 5 grams of ethyl cellulose in 150ml of dichloromethane. Then there were added 75 ml of dichloromethaneand 2.5 ml 1,2,3-propanetriol. 10 Grams of polyethylene glycol wasmolten and dissolved in 75 ml of dichloromethane. The latter solutionwas added to the former and then there were added 2.5 grams of magnesiumoctadecanoate, 5 grams of polyvinylpyrrolidone and 30 ml of concentratedcolour suspension and the whole was homogenated. The tablet cores werecoated with the thus obtained mixture in a coating apparatus.

Example E6 2% Cream

Stearyl alcohol (75 mg), cetyl alcohol (20 mg), sorbitan monostearate(20 mg) and isopropyl myristate (10 mg) are introduced in a doublewalljacketed vessel and heated until the mixture has completely molten. Thismixture is added to a seperately prepared mixture of purified water,propylene glycol (200 mg) and polysorbate 60 (15 mg) having atemperature of 70 to 75° C. while using a homogenizer for liquids. Theresulting mixture is allowed to cool to below 25° C. while continuouslymixing. A solution of A.I.(20 mg), polysorbate 80 (1 mg) and purifiedwater q.s. ad 1 g and a solution of sodium sulfite anhydrous (2 mg) inpurified water are next added to the emulsion while continuously mixing.The cream is homogenized and filled into suitable tubes.

Example E7 2% Cream

A mixture of A.I. (2 g), phosphatidyl choline (20 g), cholesterol (5 g)and ethyl alcohol (10 g) is stirred and heated at 55-60° C. untilcomplete solution and is added to a solution of methyl paraben(0.2 g),propyl paraben (0.02 g), disodium edetate (0.15 g) and sodium chloride(0.3 g) in purified water (ad 100 g) while homogenizing.Hydroxypropylmethylcellulose (1.5 g) in purified water is added and themixing is continued until swelling is complete.

What is claimed is:
 1. A compound of formula

a N-oxide form, a salt, a quaternary amine or stereochemically isomericform thereof, wherein D represents a radical of formula

wherein the dotted line represents the bond attaching D to the remainderof the compound of formula (I); X is N or CH; R³ is hydrogen or halo; R⁴is halo; —A—B— represents a bivalent radical of formula: —N═CH—  (i),—CH═N—  (ii), —CH═CH—  (iii), —CH₂—CH₂  (iv), wherein one hydrogen atomin the radicals (i) and (ii) may be replaced by a C₁₋₄alkyl radical andone or more hydrogen atoms in radicals (iii) and (iv) may be replaced bya C₁₋₄alkyl radical; Alk represents C₁₋₆alkanediyl; Y representsC₁₋₆alkanediyl optionally substituted with one or two substituentsselected from halo, hydroxy, mercapto, C₁₋₄alkyloxy, C₁₋₄alkylthio,aryloxy, arylthio, arylC₁₋₄alkyloxy, arylC₁₋₄alkylthio, cyano, amino,mono- or di(C₁₋₄alkyl)amino, mono- or di(aryl)amino, mono- ordi(arylC₁₋₄alkyl)amino, C₁₋₄alkyloxycarbonylamino,benzyloxycarbonylamino, aminocarbonyl, carboxyl, C₁₋₄alkyloxycarbonyl,guanidinyl, and aryl; R¹ represents hydrogen, C₁₋₆alkyl orarylC₁₋₆alkyl; R² represents hydrogen, C₁₋₆alkyl or arylC₁₋₆alkyl; or R¹and R² may be taken together to form a heterocyclic radical selectedfrom morpholinyl, pyrrolidinyl, piperidinyl, homopiperidinyl,piperazinyl or phthalimid-1-yl; said heterocyclic radical may optionallybe substituted with C₁₋₄alkyl, aryl, arylC₁₋₄alkyl, hydroxyC₁₋₄alkyl,amino, mono- or di(C₁₋₄alkyl)amino, aminoC₁₋₄alkyl, mono- ordi(C₁₋₄alkyl)aminoC₁₋₄alkyl, carboxyl, aminocarbonyl,C₁₋₄alkyloxycarbonyl, C₁₋₄alkyloxycarbonylamino or mono- ordi(C₁₋₄alkyl)aminocarbonyl; aryl represents phenyl, naphthalenyl,1,2,3,4-tetrahydro-naphthalenyl, indenyl or indanyl; each of said arylgroups may optionally be substituted with one or more substituentsselected from halo, C₁₋₄alkyl, hydroxy, C₁₋₄alkyloxy, nitro, amino,trifluoromethyl, hydroxyC₁₋₄alkyl, C₁₋₄alkyloxyC₁₋₄alkyl,aminoC₁₋₄alkyl, mono- or di(C₁₋₄alkyl)aminoC₁₋₄alkyl.
 2. A compoundaccording to claim 1 wherein —Alk— is a bivalent radical of formula

wherein the carbon atom marked with one asterisk is attached to thenitrogen atom and the carbon atom marked with two asterisks is attachedto the oxygen atom.
 3. A compound according to claim 1 wherein D is aradical of formula D₁ or D₂.
 4. A compound according to claim 1 whereinY is C₁₋₆alkanediyl optionally substituted with aryl.
 5. A compoundaccording to claim 1 wherein —A—B— is a radical of formula (ii).
 6. Acompound according to claim 1 which is stereoisomerically pure.
 7. Amethod for treating fungal infections in a patient, comprisingadministering an effective amount of a compound according to claim 1 tosaid patient.
 8. A pharmaceutical composition comprising apharmaceutically acceptable carrier, and as active ingredient atherapeutically effective amount of a compound as claimed in claim
 1. 9.A process of preparing a composition as claimed in claim 8 comprisingcombining a pharmaceutically acceptable carrier with a therapeuticallyeffective amount of said compound.
 10. A process of preparing a compoundaccording to claim 1, comprising a) reacting an intermediate of formula(II) with an intermediate of formula (III) in a reaction-inert solventand in the presence of a suitable base

 wherein W¹ is a suitable leaving group; b) N-alkylating an intermediateof formula (IV) with an intermediate of formula (V) in a reaction-inertsolvent in the presence of a base

 wherein W² is a suitable leaving group; and in case one or more of R1and R² is hydrogen, the primary or secondary amine group in intermediate(V) is protected with a suitable protective group P and after theN-alkylation reaction, the protected primary or secondary amine group isdeprotected to provide compounds of formula (I); c) reacting anintermediate of formula (VI) with an intermediate of formula (VII),optionally in the presence of a suitable base and optionally in areaction-inert solvent

 wherein W³ is a suitable leaving group; and in case one or more of R¹and R² is hydrogen, the primary or secondary amine group in intermediate(VII) is protected with a suitable protective group P and after theO-alkylation reaction, the protected primary or secondary amine group isdeprotected to provide compounds of formula (I).